Innate Immune Signaling in Fibrolamellar Carcinoma

纤维板层癌中的先天免疫信号传导

基本信息

批准号：
10707605
负责人：
STEEN HENNING HANSEN
金额：
$ 17.7万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-15 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10707605
关键词：
Ablation Address Affect CASP1 gene CASP8 gene CRISPR/Cas technology Carcinoma Caspase Caspase Inhibitor Catalytic Domain Cell Culture Techniques Cell Death Cells Child Chronic Cyclic AMP-Dependent Protein Kinases DNA Sequence Alteration Data Development Diagnosis Disease Disputes Engineering Exhibits Exons Extrahepatic Family Fibrolamellar Hepatocellular Carcinoma Future Gene Fusion Genes Genomics Heat shock proteins Hepatocarcinogenesis Hepatocyte IL18 gene Immune Immune signaling Immunity Infiltration Inflammasome Inflammation Inflammatory Response Injections Knock-out Link Liver Liver Stem Cell Liver neoplasms Lytic Malignant Neoplasms Malignant neoplasm of liver Mediating Modeling Mus Mutation Natural Immunity Nature Oncogenic Ontology Organoids PRKACA gene Pathogenesis Patients Phenotype Prognosis Proteins Publications Publishing Recurrence Risk Role Signal Pathway Signal Transduction Tail Testing Time Veins Work anakinra curative treatments effective therapy fusion gene genome editing hepatobiliary cancer immune cell infiltrate in vivo inhibitor insight interest liver transplantation mouse model novel preservation prevent programs stem cells tool transcriptome transcriptomic profiling transcriptomics translational study tumor tumorigenesis virtual young adult

项目摘要

Fibrolammellar Carcinoma (FLC) is a rare form of liver cancer that afflicts children and young adults. At the time of diagnosis, the cancer is stage IV in ~60% of patients without effective treatment option. For the remaining 40%, liver transplantation is the only curative therapy, providing a suitable donor can be found in time and even then, with significant risks for short- and long-term complications. Virtually all patients with FLC have a genomic alteration on Chr. 19 resulting in fusion of exon 1 of DNAJB1 of the Hsp40 family to exons 2-10 of PRKACA, a catalytic subunit of PKA. The resulting DNAJB1-PRKACA fusion gene encodes a Dnajb1-Prkaca protein with preserved catalytic activity. Using CRISPR/Cas9 technology combined with hydrodynamic tail vein injection, we and others have shown conclusively that a syntenic Dnajb1- Prkaca fusion gene on mouse Chr. 8 is sufficient to elicit liver cancer with hallmarks of FLC. FLC is thought to arise from liver stem cells, but the mechanisms whereby Dnajb1-Prkaca promotes oncogenic transformation are not well understood. To permit functional studies in a developmentally relevant context, we used CRISPR/Cas9 technology to engineer syntenic Dnajb1-Prkaca in mouse liver organoids. We discovered that liver organoids expressing Dnajb1-Prkaca are prone to undergoing lytic cell death. Next, we performed extensive transcriptome analyses that revealed an innate immune signature prominently featuring constituents of the NLRC4 inflammasome. We moreover determined that Dnajb1-Prkaca expressing liver organoids exhibit significantly elevated levels of cleaved caspase-8 consistent with its potent activation. In this work, we will first test if the NLRC4 inflammasome is activated by Dnajb1-Prkaca in mouse liver organoids and establish its role in lytic cell death. We will then determine if the Nlrc4 and/or Casp-8 genes are necessary for tumorigenesis elicited by engineering of the Dnajb1-Prkaca fusion gene in the mouse liver. The latter would represent a major breakthrough in understanding the pathogenesis of FLC.

纤维状癌癌（FLC）是一种困扰儿童和年轻人的罕见肝癌形式。当时在诊断中，癌症是约60％的没有有效治疗选择的患者的IV期。剩下的 40％的肝移植是唯一的治疗疗法，提供合适的供体可以在时间甚至可以找到合适的供体疗法然后，存在短期和长期并发症的重大风险。几乎所有FLC患者在CHR上都有基因组改变。 19导致DNAJB1外显子1的融合 HSP40家族到PRKACA 2-10的外显子，PKA的催化亚基。由此产生的DNAJB1-PRKACA融合基因编码具有保留催化活性的DNAJB1-PRKACA蛋白。使用CRISPR/CAS9技术与流体动力学的尾静脉注射结合，我们和其他人最终表明了同步DNAJB1- 小鼠chr上的prkaca融合基因。 8足以以FLC的标志引起肝癌。 FLC被认为是由肝干细胞引起的，但是DNAJB1-PRKACA促进致癌的机制转型尚不清楚。为了允许在发展相关的背景下的功能研究，我们使用CRISPR/CAS9技术来在小鼠肝脏器官中设计同时DNAJB1-PRKACA。我们发现了表达DNAJB1-PRKACA的肝癌容易遭受裂解细胞死亡。接下来，我们进行了广泛的转录组分析揭示了先天免疫签名，以选民为特征 NLRC4炎性体。此外，我们确定表达肝癌的DNAJB1-PRKACA表现裂解的caspase-8水平显着升高，与其有效激活一致。在这项工作中，我们将首先测试如果NLRC4炎症体是由小鼠肝脏类和器官中DNAJB1-PRKACA激活的并确定其在裂解细胞死亡中的作用。然后，我们将确定是否需要NLRC4和/或CASP-8基因对于小鼠肝脏中DNAJB1-PRKACA融合基因的工程引起的肿瘤发生。后者会代表了理解FLC发病机理的主要突破。